Assay for determination of daunorubicin in cancer cells with multidrug resistance phenotype
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
21641880
DOI
10.1016/j.jchromb.2011.05.008
PII: S1570-0232(11)00313-8
Knihovny.cz E-zdroje
- MeSH
- buňky K562 MeSH
- chemorezistence MeSH
- chronická myeloidní leukemie farmakoterapie metabolismus MeSH
- daunomycin analýza farmakokinetika MeSH
- DNA nádorová analýza MeSH
- fluorescence MeSH
- intracelulární prostor chemie metabolismus MeSH
- lidé MeSH
- lineární modely MeSH
- mnohočetná léková rezistence MeSH
- protinádorová antibiotika analýza farmakokinetika MeSH
- průtoková cytometrie MeSH
- senzitivita a specificita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- daunomycin MeSH
- DNA nádorová MeSH
- protinádorová antibiotika MeSH
A sensitive assay for direct determination of intracellular level of daunorubicin (DRN) in resistant leukemia cells with overexpressed P-glycoprotein has been developed. This assay is based on a rapid separation of cells from media and fast cut-off of DRN transportation by centrifugation of cells through a layer of silicone oil. Cell pellets were extracted using 1% (v/v) formic acid in 50% (v/v) ethanol in water. The cell extracts were subsequently analysed by liquid chromatography (HPLC) coupled a low-energy collision tandem mass spectrometer equipped with an electrospray ionization source (ESI-CID-MS/MS) operated in the multiple-reaction monitoring (MRM) mode. Calibration curve was linear from 0.4 to 250nM with correlation coefficient (r²) better than 0.998. The limit of quantitation (LOQ) was 0.4 nM. The assay has been successfully applied to a determination of intracellular content of daunorubicin in sensitive K562 and resistant K562/Dox and K562/HHT300 cells.
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